Page 30 - LC-SFC_Pharma_Brochure

The United States Pharmacopeia (USP) provides the following descrip-
System 1: System volume 1155 µL tion to correct for errors between systems:2) "If adjustments are neces-
6 7 sary, change in column packing (maintaining the same chemistry), the
1
2 duration of an initial isocratic hold (when prescribed), and/or dwell
3 5 volume adjustments are allowed." In other words, the adjustment of
4
initial hold time (or gradient start time adjustment) does not fall under
method change, and therefore, does not require revalidation.
System 2: System volume 505 µL (a)
2 4 : Another
3 Vendor's
System
: Nexera-i MT
1 5

System 2 + Added initial hold (+650 µL)

0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 min
ů Peaks
1 to 5: Sunscreen mixture
0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 min ů Analytical Conditions
Column : Shim-pack VP-ODS (150 mm L. × 4.6 mm I.D., 5.0 µm)
ů Peaks Mobile phase A : 20 mmol/L (Sodium) phosphate buffer solution (pH 2.5)
1. Acetophenone, 2. Propiophenone, 3. Butyrophenone, 4. Valerophenone, Mobile phase B : Acetonitrile
5. Hexanophenone, 6. Heptanophenone, 7. Octanophenone Initial concentration : 30% B
ů Analytical Conditions Flow rate : 1.0 mL/min
Column : Shim-pack XR-ODS II (75 mm L. × 3.0 mm I.D., 2.2 µm) Temp. : 60 °C
Mobile phase A : Water Detection : Photodiode array absorbance detector
Mobile phase B : Acetonitrile Injection volume : 10 µL
Initial concentration : 60% B Flow line used : HPLC Łow line (Nexera-i MT)
Flow rate : 0.8 mL/min Time program
Temp. : 40 °C Time Unit Func. Value
Detection : Photodiode array absorbance detector 2.00 Pump B.Conc 30
Injection volume : 10 µL 25.00 Pump B.Conc 70
Time program 25.10 Pump B.Conc 30
Time Unit Func. Value 32.00 Pump B.Conc 30
0.93 Pump B.Conc 60 35.00 Controller Stop
1.59 Pump B.Conc 95 Adjustment of gradient
3.50 Pump B.Conc 95
3.51 Pump B.Conc 60 start time with ACTO
6.00 Controller Stop (b)
Fig. 4 Different Retention Times Due to Different System Volumes and 2 4 : Another
Vendor's
Their Correction by Addition of an Initial Hold Time (Alkylphenones) 3 System
: Nexera-i MT
3-2. Analysis of UV Absorbents
3-2. Analysis of UV Absorbents 1 5
In this section, we describe an example method transfer to a Shimadzu
system from another vendor's HPLC system using a sunscreen mixture.
Fig. 5a shows the chromatogram obtained from the analysis of a sun-
screen mixture using Shimadzu's Nexera-i MT and another vendor's
system. Although these two analyses were performed using identical 0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 min
methods, a huge difference in the peaks after 10 min is observed. This ů Analytical Conditions (changes only)
difference is caused by the difference in system volumes of the two sys- Gradient adjustment : +340 µL (Nexera-i MT)
tems and is similar to the analysis described earlier in this report.
Retention Time Error (%) Compared to Another Vendor's System
Using ACTO's gradient start time adjustment function equipped in Component Before gradient adjustment After gradient adjustment
the Shimadzu LabSolutions workstation software, we adjusted the
gradient start time correct the difference in system volumes and per- 1 0.29 0.32
formed the analysis. As seen from Fig. 5b and the inset table in Fig. 5, 2 3 1.16 1.26
1.03
0.16
the retention times were almost identical for all peaks.
4 1.38 −0.38
Using this approach, compatibility between Shimadzu system and 5 1.46 −0.05
other vendors' system can be achieved by adjusting the gradient start
time. This means that an adjustment in the gradient start time en- Fig. 5 Example Method Transfer with Gradient Adjustment
ables smoother method transfer. (Sunscreen mixture)
3